Western blot analysis of GST tag was performed by loading various amounts of E. coli lysate containing a GST- tagged protein onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a low fluorescence PVDF membrane (Product # 22860) and blocked with SEA BLOCK (Product # 37527) for at least 1 hour. The membrane was probed with a DyLight 800-conjugated GST tag monoclonal antibody (Product # MA4-004-D800) at a dilution of 1:500 overnight at 4°C on a rocking platform and washed in TBS-0.1% Tween-20. Fluorescent detection was performed using a near IR imaging system.
|Tested species reactivity||Tag|
|Host / Isotype||Mouse / IgG|
|Immunogen||Recombinant purified full-length GST|
|Conjugate||DyLight 800 4X PEG|
|Storage buffer||PBS with proprietary stabilizer|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500-1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA4-004-D800 detects GST tag from various samples. MA4-004-D800 has been successfully used for fluorescent detection of GST tagged proteins in Western Blotting.
DyLight 800 4X PEG has an excitation/emission of 783/797nm.
Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. The GST (Glutathione-S-Transferase) is a 26kDa protein encoded by the parasite Schistosoma japonicum, easily purified by affinity chromatography on immobilized glutathione. GST is utilized as a fusion protein with foreign proteins in a range of prokaryotic expression vectors, including the pGEX family of vectors. Antibodies against GST will detect the expressed GST protein or a GST-fusion protein, and can be utilized for immuno-purification of GST-fusion proteins.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.